Mycobacterium avium subsp.

Paratuberculosis is a well established etiological agent of JD(also known as partuberculosis)

Importance
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Paratuberculosis is a chronic mycobacterial disease characterized by : irreversible wasting diarrhea and death from cachexia (wasting syndrome) in ruminants. caused by the obligate pathogen Mycobacterium avium subsp. Paratuberculosis (MAP). Subclinical carriers are estimated to produce 15-16% less milk no effective treatment. M. avium subsp. paratuberculosis has also been implicated as a possible cause of Crohns disease in humans Control programs for paratuberculosis have been established in some nations including Australia, Norway, Iceland, Japan, the Neth-erlands and the United States. (not in india)

Etiology

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Paratuberculosis results from infection by Mycobacterium avium subsp. paratuberculosis acid-fast rod previously known as Mycobacterium paratuberculosis and M. johnei Some strains of M. avium subsp. paratuberculosis preferentially infect specific hosts The two main types (distinguished by restriction fragment length polymorphisms RFLP) are the Type II or C strains, found in cattle type I or S strains, found in sheep

Species Affected

Paratuberculosis affects domesticated and wild ruminants including cattle, sheep, goats, llamas, alpaca, camels, moose, elk, bighorn sheep, buffalo, deer and reindeer. Disease has been reported in wild rabbits, nonhuman primates and pigs also.

Geographic Distribution

Paratuberculosis can be found worldwide Only Sweden and some states in Australia are proven to be free of this disease

Transmission
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In ruminants transmitted by the fecaloral route. from colostrum, milk, udder, and the male and female reproductive tracts Transmission can occur on fomites, and insects may act as mechanical vectors Young animals are most susceptible to infection Get infected when they drink milk or colostrum. In utero infections may also be seen.

M. avium subsp. paratuberculosis is resistant to environmental conditions and can survive on pastures for more than a year Viable bacteria have also been found for up to a week in bovine urine, and for up to 8-11 months in bovine feces. In one study, this organism survived for as long as nine months in pond, tap or distilled water;

Incubation Period

The incubation period is usually months or years; periods ranging from 4 months to 15 years have been reported Calves generally become infected soon after birth but rarely show clinical signs before they are two years old.

Clinical Signs
Three stages of Johnes disease in cattle: STAGE 1- Infected but showing no clinical signs and not shedding MAP. Typically this stage occurs in calves, heifers and young stock less than two years of age and many adult animals exposed to small doses of the disease-causing organism. This stage progresses slowly over many months or years to Stage II.

Stage 2- Infected, shedding MAP but showing no clinical signs. Typically this stage occurs in older heifers or adults. These animals pose a major, but often hidden, threat for infection of other animals through contamination of the environment.

Stage 3- Shedding MAP and showing clinical signs. The onset of Stage III is often associated with a period of stress, such as recent calving.
Cattle at this stage haveintermittent, watery manure. Animals lose weight and gradually drop in milk production but continue to have a good appetite. Most of these animals are shedding billions of organisms In the final and terminal aspects of Stage III of the fatal disease, animals become emaciated with fluid diarrhea and develop bottle jaw.

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Diarrhea: an early sign of Johne's disease in cattle

Signs of Johne's disease include diarrhea and rapid weight loss. Typically, affected animals remain bright and alert, without fever, and eating well.

"Bottle jaw", also called submandibular edema, is a sign very typical of Johne's disease. The cause of bottle jaw is a lack of protein in the serum, particularly albumin, due to the decreased function of the intestine. Johne's disease is a proteinlosing enteropathy.

The last portion of the small intestine, or ileum, shown in the upper part of the photo is thickened due to the inflammatory response caused by an M. paratuberculosis infection. Normally the intestine is thin and pliable as seen in the lower half of the photo. Such intestinal thickening is less pronounced in species other than cattle

Clinical signs in other animals

In sheep and goats, the wool is often damaged and easily shed, and diarrhea is less common than in cattle.

Post Mortem Lesions

In cattle the carcass may be thin or emaciated characteristic lesion is a thickened, often corrugated, wall in the distal small intestine. more advanced cases, the lesions can extend from the duodenum to the rectum. mucosa is not ulcerated mesenteric lymph nodes and other regional nodes are often enlarged and edematous. Histologicaly , the lesions are characterized by diffuse granulomatous enteritis, with the accumulation of epithelioid macrophages and giant cells in the intestinal mucosa and submucosa Acid-fast organisms may be found inside macrophages.

Enlarged mesenteric lymph nodes at necropsy

In other animals Similar lesions occur in sheep and goats The mucosa is often only slightly thickened in these species, but caseated or calcified nodules are sometimes found in the intestines and associated lymph nodes. In camelids--lymph node necrosis and mineralization can also occur.

Treatment

no satisfactory treatment some combinations of antibiotics might be successful long-term treatment would probably be required and it is not likely to be economically feasible the chance of a complete cure is low

Diagnosis
Laboratory tests variety of tests can be used . the choice of test varies with the stage of disease. subclinical carriers can be identified with serology delayedtype hypersensitivity (DTH) reactions, polymerase chain reaction (PCR) assays or fecal culture Clinical cases can be diagnosed by culturing M. avium subsp. paratuberculosis from the feces or tissues by demonstrating the organism with microscopy DNA probes or PCR

Microscopy

ZiehlNeelsen stains can be used to detect M. avium subsp. paratuberculosis in the feces

clumps of small, strongly acid-fast bacilli are diagnostic Organisms may also be found in smears from the intestinal mucosa or the cut surfaces of lymph nodes

high

power magnification photomicrograph of a ZiehlNeelsen acid-fast stained histopathology section of the bovine ileum mycobacteria (red rodshaped bacteria) can be seen

Giant cells result from fusion of several macrophages. Such cells are a hallmark of granulomatous inflammation and typical of the host response to M. paratuberculosis infection. The specific type giant cells most often seen in Johne's disease are called Langhans giant cells.

Culture
Bacteria can be cultured from the feces, thickened areas of the intestinal wall, and ileal, mesenteric and ileocecal lymph nodes Suitable media 1. Herrolds egg yolk medium 2. modified Duboss medium 3. Middlebrook 7H9 4. 7H10 and 7H11 media (mycobactin is necessary for bacterial growth) (M. avium subsp. paratuberculosis grows slowly; colonies may take 5 to 14 weeks to appear)

The standard method for diagnosis of Johne's disease is isolation of M. paratuberculosis on standard bacteriological media. Shown here is one type of such media, Herrold's egg yolk agar. Growth of slowgrowing, small, white-yellow colonies only on media containing mycobactin (two left tubes) is indicative of M. paratuberculosis. This test is usually done on fecal samples and typically takes 12 to 16 weeks.

Colonies of M. paratuberculosis grown on Middlebrook agar media without Tween are rough in appearance. Tween 80 is a surfactant commonly used in culture media to accelerate the growth of mycobacteria. For M. paratuberculosis, Tween 80 can alter the colonial and cellular morphology.

Serology and tests

Humoral immunity usually develops 10 to 17 months after infection Serology can be used for the presumptive identification of infected animals to estimate the prevalence of infection in a herd confirm paratuberculosis in animals with clinical signs (Animals that have cleared the infection can be seropositive) variety of serological tests are complement fixation enzyme-linked immunosorbent assays (ELISAs) agar gel immunodiffu-sion Intradermal testing testing with johnin or avian purified protein derivative tuberculin can detect delayed-type hypersensitiv-ity (DTH) reactions to M. avium subsp. paratuberculosis; however, this test is insensitive and nonspecific reactions are common DTH reactions may diminish or disappear as the disease progresses. Exposure to other mycobacteria, including environmental saprophytes, can result in false positives

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A simple two-day test to confirm a diagnosis of Johne's disease in cattle is the AGID serum from the animal is place in one well, a positive control is place in another, and soluble antigen is place in the third (top well). Formation of a white line of precipitation between the sample well and the antigen well (arrow) is considered a positive test. This test is commercially known as Rapid Johne's Test .

The present ELISA uses PPA antigen from the Map Bison type bacilli infecting domestic livestock population of Northern India.

Samples to collect

Although the link is speculative, veterinarians collecting samples should keep in mind that this organism may be linked to Crohns disease(a chronic enteritis of humans) M. avium subsp. paratuberculosis can be isolated from the feces, mesenteric, ileal and ileocecal lymph nodes, and thickened areas of the intestinal wall Serum samples may be taken for serology. Fresh, frozen or preserved milk samples can be tested with the milk ELISA.

The Iceberg Phenomenon infection in the herd

In the typical herd, for every animal showing clinical signs (Stage III), many other cattle are present in the earlier stages of the disease. For every obvious clinical case (Stage III) of Johnes disease on the farm, 15-25 other animals are likely infected. The clinical case represents only the tip of the iceberg of the Johnes infection.

For every stage III cow expect 1-2 more cows in Stage III (clinically diseased) 6-8 cows in Stage II (unapparent carrier adults) 10-15 cows in Stage I (infected calves & young stock)

Present scenario

Although test and removal procedures can reduce the prevalence of infection, good management practices are needed to reduce transmission within the herd There is no satisfactory treatment for the disease the trend toward intensive livestock production practices has made control more difficult Disease control is via implementing a variety of production practices described below

Management Recommendations for Farm

Calving areaConsider individual calving pens. Properly managed calving pens are ideal. Keep clean and put one cow in at a time. Focus on keeping other adult manure away from the calves. If individual calving pens are not available, be sure to keep the calving area as clean and dry as possible. Do not use calving pens for sick cows, Johnes disease positive cows or cows sick with Johnes disease. Remove calves from cows and cow area as soon after birth as possible. The sooner the better.

B. Pre-weaned heifer calves Do not feed pooled colostrum or milk. Consider pasteurization if you must feed whole milk. Limit or avoid any contact with adult cows or cow manure. Wear clean overalls & boots and use clean equipment when working with or feeding calves.

C. Post-weaned heifers Minimize any contact with adult cows or cow manure (including any manure run off that may go into heifer pens). Avoid & minimize cow manure on buckets, skid loaders, tractors, tires and other equipment.

Bred heifers Keep bred heifers separate from cows for as long as possible. Avoid feeding heifers refused (weigh back) feed from cows.

E. Cows & bulls Avoid spreading manure on hay ground or pastures, especially in the same season. Identify and manage any test-positive cows until they can be sold. Dont feed their colostrum to heifer calves.

F. Replacements & Additions Be sure to purchase replacements from herds where the Johnes status is known. Be aware of management practices and manure exposure when using a heifer raiser.

Hidden costs associated with Johnes disease include...

Premature culling of clinical or infected animals. Increased veterinary costs. Increased replacement costs. Increased susceptibility to other disease and possible breeding problems. Increase in overall cull rate. Decreased milk production in infected but normal-looking animals. (Production effect depends on parity, stage of disease and stage of lactation.) Decreased slaughter value of 20% to 30%. Decreased pool of available replacements, thereby limiting genetic pool. Decreased feed efficiency. Loss of marketing animals for sale. Loss of investment in young stock that have been infected or exposed since birth.

Job of a veterinarian
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Because Johnes disease is a hidden disease, testing should be a part of a management plan. Testing for Johnes disease can help you: Determine if an animal exhibiting defi nite clinical signs is Johnes disease positive and should be culled. Identify infected animals with suspicious clinical signs early before they further contaminate facilities and lose salvage value. Evaluate the extent of infection in your herd. Monitor progress of control efforts. Know if you are marketing infected or low-risk cattle and, as a result, know if you are contributing to the spread of the disease to producers herds or helping producers prevent Johnes disease from entering their herd. Know if you are about to purchase a Johnes disease test-positive or low-risk animal before its brought into the herd.

Following table contents can be followed by the veterinarian to control JD in a herd